CN106432437A - Antigen protein of bovine respiration syncytial viruses - Google Patents

Antigen protein of bovine respiration syncytial viruses Download PDF

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Publication number
CN106432437A
CN106432437A CN201611065849.2A CN201611065849A CN106432437A CN 106432437 A CN106432437 A CN 106432437A CN 201611065849 A CN201611065849 A CN 201611065849A CN 106432437 A CN106432437 A CN 106432437A
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ser
val
leu
asn
ile
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汤斌
闫波
姜钧耀
汤明
孟祥斌
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Yantai Royal Emperor Biological Engineering Co Ltd
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Yantai Royal Emperor Biological Engineering Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2299/00—Coordinates from 3D structures of peptides, e.g. proteins or enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18511—Pneumovirus, e.g. human respiratory syncytial virus
    • C12N2760/18522—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

The invention discloses an antigen protein of bovine respiration syncytial viruses. At least two sites of an amino acid sequence of the antigen protein of the bovine respiration syncytial viruses are substituted by cysteine as compared with SEQ CAA76980.1, the substituted cysteine is connected by disulfide bonds, and internal disulfide bonds, hole filling, single-strain connection and the like are improved on the basis. The invention further provides a method for preparing the antigen protein. The antigen protein and the method have the advantage that technologies and theories of protein crystallography are applied to the antigen protein, structural change of the antigen protein in virus infection and pathogenicity procedures can be determined, genetic engineering modification can be carried out on critical virus proteins according to results of the structural change, accordingly, the virus proteins can become protein antigens with a function of infection without pathogenicity and capability of causing efficient reaction of antibodies of animal bodies, and bovine respiration syncytial virus infection can be effectively prevented and treated; perfect virus vaccine can be developed on the basis, accordingly, loss due to virus infection can be effectively controlled and prevented, and the antigen protein and the method have important economical significance and social significance.

Description

A kind of bovine respiratory born of the same parents zoarium viral antigen proteins
Technical field
The invention belongs to biological product preparing technical field, be related to a kind of bovine respiratory born of the same parents zoarium viral antigen proteins and its Preparation method.
Background technology
The fit virus of bovine respiratory born of the same parents（Bovine Respiration Syncytial Virus – BRSV）Infection can be drawn Cattle respiratory disease is played, is listed in EU member country and is only second to bovine mucosal disease（BVD）And infectious bovine rhinotrachetiss（IBR）'s One of important cattle disease.Primary disease is popular in world wide, and its hazardness is that BRSV the incidence of infection is up to 60%-80%, extremely Rate of dying reaches more than 20% in some outbursts, causes very big economic loss to cattle-raising.Although this BRSV is from eighties of last century Begin to the seventies list research and development category in, do not develop preferable medicine and vaccine for various reasons.Although relevant In multiple inactivations of BRSV, weak poison, the report of restructuring and DNA vaccination, but there is certain distance apart from Clinical practice, current state Still without preferable viral vaccine on border.
The development of traditional vaccine and production are mainly by changing condition of culture, or pass on different host animals and make cause Sick Ecotoxicology weakens, or inactivated to complete by physics, chemical method.With the continuous progress of human knowledge, The limitation of traditional vaccine is also increasingly revealed：(1) virus of animals and humans needs to cultivate in zooblast, and this causes The cost of production of vaccine is very high；(2) morbid substance in vaccine is possible in vaccine production process not kill completely or fill Divide attenuation, this can cause containing strong toxicity morbid substance in vaccine, and then disease is propagated in the larger context；(3) subtract Toxic bacterial strain is possible to undergo mutation, and causes disease.
The open beginning for having created Chinese structure Basic of Biology theoretical research is applied, structure biology is applied to by the present invention In practice, synthesis bovine respiratory born of the same parents zoarium viral antigen proteins.Bovine respiratory born of the same parents zoarium virus is family of paramyxovirus section Member, F- albumen thereon is highly conserved and forms trimer furcella, which occurs conformation change under activation.F- albumen is situated between The fusion of virus and cell membrane is led, allows viral nucleocapsid that Cytoplasm is entered, the step for suppressing F- protein mediated prevents infectious cycle Initial period and neutralize viral infection and can effectively prevent virus infraction, i.e., for the antibody of F- albumen, suppress its Activity, can neutralize viral infection and can be protected from BRSV infection.Apply a disclosure and create Chinese structure Basic of Biology The beginning of theoretical research, structure biology is applied in practice by the present invention, synthesis bovine respiratory born of the same parents' zoarium viral antigen proteins.
Content of the invention
It is an object of the invention to the shortcoming for overcoming prior art to exist, provides a kind of bovine respiratory born of the same parents zoarium virus antigen Albumen and preparation method thereof, the technology of the antigen protein application protein crystallography and theory, determine albumen in virus infection and Structure change in pathogenic course, the result further according to structure change carries out genetic engineering to this crucial virus protein and changes Make so as to become and only infect proteantigen that is not pathogenic and causing animal body antibody highly effective reaction.
To achieve these goals, the present invention solves its technical problem and is adopted the technical scheme that：
A kind of bovine respiratory born of the same parents zoarium viral antigen proteins, the fit virus key protein F protein wild species of coding bovine respiratory born of the same parents Aminoacid sequence abbreviation SEQ CAA76980.1, particular sequence is MATTAMTMIISIIFISTYVTHITLCQNITEEFYQSTCSAVSRGYLSALRTGWYTSVVTIELSKIQKNVCKSTDSKVK LIKQELERYNNAVVELQSLMQNEPASFSRAKRSIPELIHYTRNSTKKFYGLMGKKRKRRFLGFLLGIGSAIASGVAV SKVLHLEGEVNKIKNALLSTNKAVVSLSNGVSVLTSKVLDLKNYIDKELLPKVNNHDCRISNIATVIEFQQKNNRLL EIAREFSVNAGITTPLSTYMLTNSELLSLINDMPITNDQKKLMSSNVQIVRQQSYSIMSVVKEEVIAYVVQLPIYGV IDTPCWKLHTSPLCTTDNKEGSNICLTRTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPTDVNLCNTD IFNTKYDCKIMTSKTDISSSVITSIGAIVCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTLYYVNKLE GKALYIKGEPIINYYDPLVFPSDEFDASIAQVNAKINQSLAFIRRSDELLHSVDVGKSTTNVVITTIIIVIVVVILM LIAVGLLFYCKTRSTPIMLGKDQLSGINNLSFSK, it is characterised in that：The bovine respiratory born of the same parents zoarium viral antigen proteins Aminoacid sequence compare at least two sites of SEQ CAA76980.1 and substituted by cysteine, and half Guang ammonia after substituting By disulfide bond between acid.
Used as optimized, the site is the 148th L-Valine of aminoacid sequence and 288 isoleucine.
Used as optimized, the site is the 158th Leucine of aminoacid sequence and 290 serines.
Used as optimized, the aminoacid sequence of the bovine respiratory born of the same parents zoarium viral antigen proteins compares SEQ CAA76980.1, is substituted by cysteine in the Leucine in the 260th site, and realizes hole filling in the site.
Used as optimized, the aminoacid sequence for bovine respiratory born of the same parents zoarium viral antigen proteins rejects SEQ The aminoacid sequence in 103 sites in CAA76980.1 sequence to 138 sites, connects 102 sites and 139 using sgsgs Point.
Used as optimized, the aminoacid sequence for bovine respiratory born of the same parents zoarium viral antigen proteins rejects SEQ The aminoacid sequence in 103 sites in CAA76980.1 sequence to 142 sites, connects 102 sites and 143 using sgsgs Point.
Used as optimized, the aminoacid sequence of the coding bovine respiratory born of the same parents zoarium viral antigen proteins compares SEQ CAA76980.1 sequence, the 99th site aspartic acid is substituted by cysteine, and the 362nd site serine is substituted by cysteine, And between the cysteine after replacement connect in disulfide bond.
A kind of preparation method of bovine respiratory born of the same parents zoarium viral antigen proteins, it is characterised in that：Concrete technology is as follows：
（1）According to the principle of structure biology, protein crystal is carried out, observed with X-ray after crystallization, determine and infect virus key Cause of disease F protein infect before and after structure change；
（2）According to step（1）Structure change designs the sequence of antigen protein and structure；
（3）Determine and need the qualification of antigen protein through multiple antigen tests and stability characteristic detection；
（4）Antigen protein sequence through checking is carried out corresponding albumen synthesis, expression, purification and is detected, obtain for making For animal experiment and clinical trial for bovine respiratory born of the same parents zoarium viral antigen proteins.
The protein mediated virus of cattle born of the same parents zoarium virus key F- and the fusion of cell membrane, allow viral nucleocapsid enter cell.Pin Antigen protein to bovine respiratory born of the same parents zoarium viral design, is that the structure change before and after being infected according to F- albumen sets, State before making F protein structure remain at fusion on the basis of genetic engineering so as to only infecting not pathogenic and cause The features such as proteantigen of animal body antibody highly effective reaction.
The present invention can make F protein keep trimer furcella state before and after infecting（Closure state）, only infected Proteantigen that is not pathogenic and causing animal's antibody highly effective reaction；In order to ensure that coming from the F- albumen on pathogenic virus is in Stable preinfective state, to the connects chain for adding series on F- albumen, including interior disulfide bond, disulfide bond, hole filling With single-stranded connection.
Compared with prior art, its advantage is the present invention：The bovine respiratory for being prepared using protein crystallography method Born of the same parents' zoarium viral antigen proteins can effectively prevent and treat the fit virus infection of bovine respiratory born of the same parents, can open based on this Send preferable viral vaccine；The loss that effective control and strick precaution virus infection bring, with great economic implications and society Meaning.
Description of the drawings
Fig. 1 the present invention relates to the use of Phenix software and carry out the 3-D solid structure simulation of cattle syncytial viruses F- albumen Figure, left figure is trimer 3-D solid structure simulation drawing before contaminating, and right figure is the partial enlarged drawing of left figure lower right corner square frame position.
Fig. 2 cattle syncytial viruses Respiroviruses according to the present invention infect the F- protein structure of the key of before and after's process Change schematic diagram.Before in figure shows and infects（Left）, F protein trimer assumes trimer furcella state, after infecting（Right）F- egg It is activated in vain, occurred conformation changes, the fusion of virus and cell membrane, viral nucleocapsid enters Cytoplasm, and then infects cattle body simultaneously So which is caused a disease.
Fig. 3 Electronic Speculum for infecting Niu Tihou through the successful bovine respiratory born of the same parents zoarium proteantigen of transformation according to the present invention Photo state.Left figure shows that the bovine respiratory born of the same parents zoarium proteantigen of RD-BRSV-DB1-CF1 transformation infects the state of Niu Tihou, It can be seen that assuming trimer furcella state before stable fusion, right figure is the bovine respiratory born of the same parents zoarium virus infection cattle without transformation Electromicroscopic photograph after body.
Add connects chain schematic diagram on Fig. 4 F- albumen according to the present invention, including interior disulfide bond, disulfide bond, hole filling With single-stranded connection, these links guarantee F- albumen holding trimer furcella state.
Specific embodiment
Below by embodiment and combine accompanying drawing the present invention is further described.
Embodiment 1：SEQ CAA76980.1 is carried out a kind of bovine respiratory born of the same parents zoarium disease that following transformation obtains the present invention Malicious antigen protein：
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 148th L-Valine of CAA76980.1 is replaced by cysteine In generation, the 288th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond, is carried out corresponding albumen conjunction Become, express, purify, the bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB1 for obtaining, its aminoacid sequence is referred to as For SEQ DB1.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 158th Leucine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 290th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond, is carried out corresponding egg White synthesis, expression, purification, the bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB2 for obtaining, its aminoacid sequence Referred to as SEQ DB2.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 148th L-Valine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 288th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond, while by the 260th site Leucine is substituted by cysteine, and realizes hole filling in the site, is carried out corresponding albumen synthesis, expression, purification, The bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB1-CF1 for obtaining, its aminoacid sequence is referred to as SEQ DB1-CF1.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 158th Leucine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 290th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond, while by the 260th site Leucine is substituted by cysteine, and realizes hole filling in the site, is carried out corresponding albumen synthesis, expression, purification, The bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB2-CF1 for obtaining, its aminoacid sequence is referred to as SEQ DB2-CF1.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 148th L-Valine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 288th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond；Simultaneously by the 260th site Leucine is substituted by cysteine, and realizes hole filling in the site；Reject 103 to position in SEQ CAA76980.1 sequence Point 138 sites aminoacid sequence, using sgsgs connect 102 and 139 sites, carried out corresponding albumen synthesis, expression, Purification, the bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB1-CF1-SC1 for obtaining, its aminoacid sequence is referred to as For SEQ DB1-CF1-SC1.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 158th Leucine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 290th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond, while by the 260th site Leucine is substituted by cysteine, and realizes hole filling in the site；Reject 103 to position in SEQ CAA76980.1 sequence Point 138 sites aminoacid sequence, using sgsgs connect 102 and 139 sites, carried out corresponding albumen synthesis, expression, Purification, the bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB2-CF1-SC1 for obtaining, its aminoacid sequence is referred to as For SEQ DB2-CF1-SC1.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 148th L-Valine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 288th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond；Simultaneously by the 260th site Leucine is substituted by cysteine, and realizes hole filling in the site；Reject 103 to position in SEQ CAA76980.1 sequence The aminoacid sequence in 138 sites of point, connects 102 and 139 sites using sgsgs, and the 99th site aspartic acid is replaced by cysteine In generation, the 362nd site serine is substituted by cysteine, and disulfide bond in connecting between the cysteine after replacement, carries out phase The albumen that answers synthesizes, expresses, purification, the bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB1-CF1- for obtaining SC1-ID1, its aminoacid sequence is referred to as SEQ DB1-CF1-SC1-ID1.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 148th L-Valine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 288th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond；Simultaneously by the 260th site Leucine is substituted by cysteine, and realizes hole filling in the site；Reject 103 to position in SEQ CAA76980.1 sequence The aminoacid sequence in 142 sites of point, connects 102 and 143 sites using sgsgs, and the 99th site aspartic acid is replaced by cysteine In generation, the 362nd site serine is substituted by cysteine, and disulfide bond in connecting between the cysteine after replacement, carries out phase The albumen that answers synthesizes, expresses, purification, the bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB1-CF1- for obtaining SC2-ID1, its aminoacid sequence is referred to as SEQ DB1-CF1-SC2-ID1.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 158th Leucine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 290th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond, while by the 260th site Leucine is substituted by cysteine, and realizes hole filling in the site；Reject 103 to position in SEQ CAA76980.1 sequence The aminoacid sequence in 138 sites of point, connects 102 and 139 sites using sgsgs；99th site aspartic acid is replaced by cysteine In generation, the 362nd site serine is substituted by cysteine, and disulfide bond in connecting between the cysteine after replacement.It carries out Corresponding albumen synthesis, expression, purification, the bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB2-CF1- for obtaining SC1-ID1, its aminoacid sequence is referred to as SEQ DB2-CF1-SC1-ID1.
Bovine respiratory born of the same parents zoarium virus F protein aminoacid sequence SEQ the 158th Leucine of CAA76980.1 is by half Guang ammonia Acid is substituted, and the 290th isoleucine is substituted by cysteine, and is connected two sites by disulfide bond, while by the 260th site Leucine is substituted by cysteine, and realizes hole filling in the site；Reject 103 to position in SEQ CAA76980.1 sequence The aminoacid sequence in 142 sites of point, connects 102 and 143 sites using sgsgs；99th site aspartic acid is replaced by cysteine In generation, the 362nd site serine is substituted by cysteine, and disulfide bond in connecting between the cysteine after replacement.It carries out Corresponding albumen synthesis, expression, purification, the bovine respiratory born of the same parents zoarium viral antigen proteins abbreviation RD-BRSV-DB2-CF1- for obtaining SC2-ID1, its aminoacid sequence is referred to as SEQ DB2-CF1-SC2-ID1.
Also known as antigenicity, immunogenicity refers to that antigenic stimulus body produces the characteristic of immunne response ability.Immunoreactivity refers to Antigen molecule can be with the product of corresponding immunne response（Antibody or primed lymphocyte）, specifically bind in vivo or in vitro Performance, the affinity-immunity using ELISA method to antigen with antibody tests.ELISA method has sensitive, special, simple Singly, quickly the features such as, stablizing and be easily operated automatically, as a new technique in immunologic diagnosises, is applied successfully to Macromole antigen and the quantitative determination of small molecule antigens.The antibody of high affinity and the adhesion of antigen are strong, i.e., antigen concentration is very Also more antibodies bind antigen forms immune complex when low.Affinity is represented with equilibrium constant K, and K value is bigger, affine Property is higher, also more firm with antigen binding.Table 1 give the present invention preparation antigen protein respectively Site, SiteQP, The affinity data of Site V, II site of Site and antibodies.
The Characteristics Detection of 1 embodiment bovine respiratory born of the same parents of table zoarium viral antigen proteins
The present invention has higher antibody library affinity through the F- albumen of design improvement.All protect through improved antigen protein Hold trimer furcella state before fusion（Trimer）, and purpose antigen albumen can be obtained.
The present invention is obtained the antigen protein of purification and is specifically bound with D25 antibody, through protein immunization electrophoresis and ELISA Method is detected, as a result shows not only there is preferable immunogenicity, and can inducing mouse produce higher antibody horizontal, table 2 give the immunogenicity weighted mean that D25 antibody is specifically bound with antigen protein of the present invention, and matched group is unmanifest open country Non-hibernating eggs.
The Characteristics Detection that 2 embodiment bovine respiratory born of the same parents of table zoarium viral antigen proteins are specifically bound with D25
Antigen protein D25 affinity potency weighted mean（）
RD-BRSV-DB1 4659
RD-BRSV-DB2 547
RD-BRSV-DB1-CF1 4756
RD-BRSV-DB2-CF1 875
RD-BRSV-DB1-CF1-SC1 3689
RD-BRSV-DB2-CF1-SC1 4549
RD-BRSV-DB1-CF1-SC1-ID1 10568
RD-BRSV-DB1-CF1-SC2-ID1 1534
RD-BRSV-DB2-CF1-SC1-ID1 1895
RD-BRSV-DB2-CF1-SC2-ID1 5668
Unmanifest wild species 100
Physical and chemical stability is carried out to the antigen protein in the present invention to be measured, including aspects such as temperature, pH, osmotic pressuries, Detailed data is shown in Table 3, and in table, data are the active ratio with optimal antigen protein activity of antigen protein under corresponding conditionses, knot Fruit shows to assume preferable stability through the antigen protein of design improvement to temperature, pH value, osmotic pressure etc., with higher anti- Should activity.
The reactivity stability of 3 embodiment bovine respiratory born of the same parents of table zoarium viral antigen proteins
Embodiment 2：Prepare the concrete technology of RD-BRSV-DB1-CF1-SC1-ID1 in embodiment 1.See albumen mechanism and shape Can be by following three kinds of approach, respectively x- ray（x-ray Crystallography）, nuclear magnetic resonance, NMR（NMR）, Electronic Speculum, The present invention is observed to protein structure using x-ray method.Serial by target egg using 600 plux of Solution maker Crystallized in vain, the target protein after crystallization is adopted X-ray system（SER-CAT22）Crystallization volume data is collected, and becomes which Picture, obtains the electronic cloud of protein structure, obtains the data variation before and after key protein F- albumen infects, and infects front-end geometry change Change as shown in Figure 2.
Data processing of racking is entered to above-mentioned electronic cloud, using corresponding software（Phenix）Carry out with wild protein sequence Digital simulation, forms the 3-D solid structure simulation figure of protein structure.According to the three-dimensional protein structure that protein sequence is simulated Figure obtains the Trimeric structures of F- albumen, refers to Fig. 1.
According to the principle of crystal chemistry, antigen protein structure design is carried out using simulation softward, its design is included as Fig. 4 institute The four kinds of modes that shows：Interior disulfide bond（intra disulphide bond）, disulfide bond（disulphide bond）, hole filling （cavity filling）With single-stranded connection（single chain）, wild species CAA76980.1 is designed.
The protein sequence for designing is translated into DNA sequence, the Expi293F conduct for providing using invitrogen company Cell carrier carries out in 293Fectin culture medium cultivating 5 days, collects cell culture fluid, target protein is carried out purification Obtain the BRSV F- albumen of purification for designing, i.e. RD-BRSV-DB1-CF1-SC1-ID1, improved bovine respiratory born of the same parents close Body protein antigen infects trimer furcella state before Niu Tihou assumes stable fusion and sees Fig. 3（Left）.
Above-mentioned specific embodiment is only the concrete case of the present invention, and the scope of patent protection of the present invention is included but is not limited to The product form of above-mentioned specific embodiment and style, a kind of any bovine respiratory born of the same parents zoarium for meeting claims of the present invention Appropriate change or modification that viral antigen proteins and any person of an ordinary skill in the technical field are done to which, should all fall into The scope of patent protection of the present invention.
SEQUENCE LISTING
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<120>A kind of bovine respiratory born of the same parents zoarium viral antigen proteins
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Val Ser Lys Val Leu His Leu Glu Gly Glu Val Asn Lys Ile Lys Asn
130 135 140
Ala Leu Leu Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly Val
145 150 155 160
Ser Val Leu Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp Lys
165 170 175
Glu Leu Leu Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn Ile
180 185 190
Ala Thr Val Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu Ile
195 200 205
Ala Arg Glu Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser Thr
210 215 220
Tyr Met Leu Thr Asn Ser Glu Leu Leu Ser Leu Ile Asn Asp Met Pro
225 230 235 240
Ile Thr Asn Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile Val
245 250 255
Arg Gln Gln Ser Tyr Ser Cys Met Ser Val Val Lys Glu Glu Val Ile
260 265 270
Ala Tyr Val Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro Cys
275 280 285
Trp Lys Leu His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu Gly
290 295 300
Ser Asn Ile Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp Asn
305 310 315 320
Ala Gly Ser Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val Gln
325 330 335
Ser Asn Arg Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro Thr
340 345 350
Asp Val Asn Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp Cys
355 360 365
Lys Ile Met Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr Ser
370 375 380
Ile Gly Ala Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser Asn
385 390 395 400
Lys Asn Arg Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr Val
405 410 415
Ser Asn Lys Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr Tyr
420 425 430
Val Asn Lys Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro Ile
435 440 445
Ile Asn Tyr Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp Ala
450 455 460
Ser Ile Ala Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe Ile
465 470 475 480
Arg Arg Ser Asp Glu Leu Leu His
485
<210> 3
<211> 488
<212> PRT
<213>Artificial sequence
<400> 3
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Asn Glu Pro Ala Ser Phe Ser
65 70 75 80
Arg Ala Lys Arg Ser Ile Pro Glu Leu Ile His Tyr Thr Arg Asn Ser
85 90 95
Thr Lys Lys Phe Tyr Gly Leu Met Gly Lys Lys Arg Lys Arg Arg Phe
100 105 110
Leu Gly Phe Leu Leu Gly Ile Gly Ser Ala Ile Ala Ser Gly Val Ala
115 120 125
Val Ser Lys Val Cys His Leu Glu Gly Glu Val Asn Lys Ile Lys Asn
130 135 140
Ala Leu Leu Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly Val
145 150 155 160
Ser Val Leu Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp Lys
165 170 175
Glu Leu Leu Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn Ile
180 185 190
Ala Thr Val Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu Ile
195 200 205
Ala Arg Glu Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser Thr
210 215 220
Tyr Met Leu Thr Asn Ser Glu Leu Leu Ser Leu Ile Asn Asp Met Pro
225 230 235 240
Ile Thr Asn Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile Val
245 250 255
Arg Gln Gln Ser Tyr Ser Ile Met Cys Val Val Lys Glu Glu Val Ile
260 265 270
Ala Tyr Val Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro Cys
275 280 285
Trp Lys Leu His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu Gly
290 295 300
Ser Asn Ile Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp Asn
305 310 315 320
Ala Gly Ser Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val Gln
325 330 335
Ser Asn Arg Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro Thr
340 345 350
Asp Val Asn Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp Cys
355 360 365
Lys Ile Met Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr Ser
370 375 380
Ile Gly Ala Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser Asn
385 390 395 400
Lys Asn Arg Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr Val
405 410 415
Ser Asn Lys Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr Tyr
420 425 430
Val Asn Lys Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro Ile
435 440 445
Ile Asn Tyr Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp Ala
450 455 460
Ser Ile Ala Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe Ile
465 470 475 480
Arg Arg Ser Asp Glu Leu Leu His
485
<210> 4
<211> 488
<212> PRT
<213>Artificial sequence
<400> 4
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Asn Glu Pro Ala Ser Phe Ser
65 70 75 80
Arg Ala Lys Arg Ser Ile Pro Glu Leu Ile His Tyr Thr Arg Asn Ser
85 90 95
Thr Lys Lys Phe Tyr Gly Leu Met Gly Lys Lys Arg Lys Arg Arg Phe
100 105 110
Leu Gly Phe Leu Leu Gly Ile Gly Ser Ala Cys Ala Ser Gly Val Ala
115 120 125
Val Ser Lys Val Leu His Leu Glu Gly Glu Val Asn Lys Ile Lys Asn
130 135 140
Ala Leu Leu Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly Val
145 150 155 160
Ser Val Leu Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp Lys
165 170 175
Glu Leu Leu Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn Ile
180 185 190
Ala Thr Val Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu Ile
195 200 205
Ala Arg Glu Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser Thr
210 215 220
Tyr Met Leu Thr Asn Ser Glu Leu Leu Ser Phe Ile Asn Asp Met Pro
225 230 235 240
Ile Thr Asn Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile Val
245 250 255
Arg Gln Gln Ser Tyr Ser Cys Met Ser Val Val Lys Glu Glu Val Ile
260 265 270
Ala Tyr Val Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro Cys
275 280 285
Trp Lys Leu His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu Gly
290 295 300
Ser Asn Ile Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp Asn
305 310 315 320
Ala Gly Ser Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val Gln
325 330 335
Ser Asn Arg Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro Thr
340 345 350
Asp Val Asn Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp Cys
355 360 365
Lys Ile Met Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr Ser
370 375 380
Ile Gly Ala Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser Asn
385 390 395 400
Lys Asn Arg Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr Val
405 410 415
Ser Asn Lys Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr Tyr
420 425 430
Val Asn Lys Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro Ile
435 440 445
Ile Asn Tyr Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp Ala
450 455 460
Ser Ile Ala Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe Ile
465 470 475 480
Arg Arg Ser Asp Glu Leu Leu His
485
<210> 5
<211> 488
<212> PRT
<213>Artificial sequence
<400> 5
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Asn Glu Pro Ala Ser Phe Ser
65 70 75 80
Arg Ala Lys Arg Ser Ile Pro Glu Leu Ile His Tyr Thr Arg Asn Ser
85 90 95
Thr Lys Lys Phe Tyr Gly Leu Met Gly Lys Lys Arg Lys Arg Arg Phe
100 105 110
Leu Gly Phe Leu Leu Gly Ile Gly Ser Ala Ile Ala Ser Gly Val Ala
115 120 125
Val Ser Lys Val Cys His Leu Glu Gly Glu Val Asn Lys Ile Lys Asn
130 135 140
Ala Leu Leu Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly Val
145 150 155 160
Ser Val Leu Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp Lys
165 170 175
Glu Leu Leu Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn Ile
180 185 190
Ala Thr Val Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu Ile
195 200 205
Ala Arg Glu Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser Thr
210 215 220
Tyr Met Leu Thr Asn Ser Glu Leu Leu Ser Phe Ile Asn Asp Met Pro
225 230 235 240
Ile Thr Asn Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile Val
245 250 255
Arg Gln Gln Ser Tyr Ser Ile Met Cys Val Val Lys Glu Glu Val Ile
260 265 270
Ala Tyr Val Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro Cys
275 280 285
Trp Lys Leu His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu Gly
290 295 300
Ser Asn Ile Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp Asn
305 310 315 320
Ala Gly Ser Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val Gln
325 330 335
Ser Asn Arg Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro Thr
340 345 350
Asp Val Asn Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp Cys
355 360 365
Lys Ile Met Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr Ser
370 375 380
Ile Gly Ala Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser Asn
385 390 395 400
Lys Asn Arg Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr Val
405 410 415
Ser Asn Lys Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr Tyr
420 425 430
Val Asn Lys Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro Ile
435 440 445
Ile Asn Tyr Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp Ala
450 455 460
Ser Ile Ala Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe Ile
465 470 475 480
Arg Arg Ser Asp Glu Leu Leu His
485
<210> 6
<211> 457
<212> PRT
<213>Artificial sequence
<400> 6
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Asn Glu Pro Ala Ser Gly Ser
65 70 75 80
Gly Ser Gly Phe Leu Leu Gly Ile Gly Ser Ala Cys Ala Ser Gly Val
85 90 95
Ala Val Ser Lys Val Leu His Leu Glu Gly Glu Val Asn Lys Ile Lys
100 105 110
Asn Ala Leu Leu Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly
115 120 125
Val Ser Val Leu Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp
130 135 140
Lys Glu Leu Leu Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn
145 150 155 160
Ile Ala Thr Val Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu
165 170 175
Ile Ala Arg Glu Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser
180 185 190
Thr Tyr Met Leu Thr Asn Ser Glu Leu Leu Ser Phe Ile Asn Asp Met
195 200 205
Pro Ile Thr Asn Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile
210 215 220
Val Arg Gln Gln Ser Tyr Ser Cys Met Ser Val Val Lys Glu Glu Val
225 230 235 240
Ile Ala Tyr Val Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro
245 250 255
Cys Trp Lys Leu His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu
260 265 270
Gly Ser Asn Ile Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp
275 280 285
Asn Ala Gly Ser Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val
290 295 300
Gln Ser Asn Arg Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro
305 310 315 320
Thr Asp Val Asn Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp
325 330 335
Cys Lys Ile Met Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr
340 345 350
Ser Ile Gly Ala Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser
355 360 365
Asn Lys Asn Arg Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr
370 375 380
Val Ser Asn Lys Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr
385 390 395 400
Tyr Val Asn Lys Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro
405 410 415
Ile Ile Asn Tyr Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp
420 425 430
Ala Ser Ile Ala Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe
435 440 445
Ile Arg Arg Ser Asp Glu Leu Leu His
450 455
<210> 7
<211> 453
<212> PRT
<213>Artificial sequence
<400> 7
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Asn Glu Pro Ala Ser Gly Ser
65 70 75 80
Gly Ser Gly Ile Gly Ser Ala Ile Ala Ser Gly Val Ala Val Ser Lys
85 90 95
Val Cys His Leu Glu Gly Glu Val Asn Lys Ile Lys Asn Ala Leu Leu
100 105 110
Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly Val Ser Val Leu
115 120 125
Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp Lys Glu Leu Leu
130 135 140
Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn Ile Ala Thr Val
145 150 155 160
Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu Ile Ala Arg Glu
165 170 175
Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser Thr Tyr Met Leu
180 185 190
Thr Asn Ser Glu Leu Leu Ser Phe Ile Asn Asp Met Pro Ile Thr Asn
195 200 205
Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile Val Arg Gln Gln
210 215 220
Ser Tyr Ser Ile Met Cys Val Val Lys Glu Glu Val Ile Ala Tyr Val
225 230 235 240
Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro Cys Trp Lys Leu
245 250 255
His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu Gly Ser Asn Ile
260 265 270
Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp Asn Ala Gly Ser
275 280 285
Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val Gln Ser Asn Arg
290 295 300
Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro Thr Asp Val Asn
305 310 315 320
Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp Cys Lys Ile Met
325 330 335
Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr Ser Ile Gly Ala
340 345 350
Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser Asn Lys Asn Arg
355 360 365
Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr Val Ser Asn Lys
370 375 380
Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr Tyr Val Asn Lys
385 390 395 400
Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro Ile Ile Asn Tyr
405 410 415
Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp Ala Ser Ile Ala
420 425 430
Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe Ile Arg Arg Ser
435 440 445
Asp Glu Leu Leu His
450
<210> 8
<211> 457
<212> PRT
<213>Artificial sequence
<400> 8
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Cys Glu Pro Ala Ser Gly Ser
65 70 75 80
Gly Ser Gly Phe Leu Leu Gly Ile Gly Ser Ala Cys Ala Ser Gly Val
85 90 95
Ala Val Ser Lys Val Leu His Leu Glu Gly Glu Val Asn Lys Ile Lys
100 105 110
Asn Ala Leu Leu Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly
115 120 125
Val Ser Val Leu Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp
130 135 140
Lys Glu Leu Leu Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn
145 150 155 160
Ile Ala Thr Val Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu
165 170 175
Ile Ala Arg Glu Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser
180 185 190
Thr Tyr Met Leu Thr Asn Ser Glu Leu Leu Ser Phe Ile Asn Asp Met
195 200 205
Pro Ile Thr Asn Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile
210 215 220
Val Arg Gln Gln Ser Tyr Ser Cys Met Ser Val Val Lys Glu Glu Val
225 230 235 240
Ile Ala Tyr Val Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro
245 250 255
Cys Trp Lys Leu His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu
260 265 270
Gly Ser Asn Ile Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp
275 280 285
Asn Ala Gly Ser Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val
290 295 300
Gln Cys Asn Arg Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro
305 310 315 320
Thr Asp Val Asn Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp
325 330 335
Cys Lys Ile Met Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr
340 345 350
Ser Ile Gly Ala Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser
355 360 365
Asn Lys Asn Arg Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr
370 375 380
Val Ser Asn Lys Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr
385 390 395 400
Tyr Val Asn Lys Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro
405 410 415
Ile Ile Asn Tyr Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp
420 425 430
Ala Ser Ile Ala Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe
435 440 445
Ile Arg Arg Ser Asp Glu Leu Leu His
450 455
<210> 9
<211> 453
<212> PRT
<213>Artificial sequence
<400> 9
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Cys Glu Pro Ala Ser Gly Ser
65 70 75 80
Gly Ser Gly Ile Gly Ser Ala Ile Ala Ser Gly Val Ala Val Ser Lys
85 90 95
Val Cys His Leu Glu Gly Glu Val Asn Lys Ile Lys Asn Ala Leu Leu
100 105 110
Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly Val Ser Val Leu
115 120 125
Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp Lys Glu Leu Leu
130 135 140
Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn Ile Ala Thr Val
145 150 155 160
Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu Ile Ala Arg Glu
165 170 175
Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser Thr Tyr Met Leu
180 185 190
Thr Asn Ser Glu Leu Leu Ser Phe Ile Asn Asp Met Pro Ile Thr Asn
195 200 205
Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile Val Arg Gln Gln
210 215 220
Ser Tyr Ser Ile Met Cys Val Val Lys Glu Glu Val Ile Ala Tyr Val
225 230 235 240
Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro Cys Trp Lys Leu
245 250 255
His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu Gly Ser Asn Ile
260 265 270
Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp Asn Ala Gly Ser
275 280 285
Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val Gln Cys Asn Arg
290 295 300
Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro Thr Asp Val Asn
305 310 315 320
Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp Cys Lys Ile Met
325 330 335
Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr Ser Ile Gly Ala
340 345 350
Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser Asn Lys Asn Arg
355 360 365
Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr Val Ser Asn Lys
370 375 380
Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr Tyr Val Asn Lys
385 390 395 400
Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro Ile Ile Asn Tyr
405 410 415
Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp Ala Ser Ile Ala
420 425 430
Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe Ile Arg Arg Ser
435 440 445
Asp Glu Leu Leu His
450
<210> 10
<211> 457
<212> PRT
<213>Artificial sequence
<400> 10
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Cys Glu Pro Ala Ser Gly Ser
65 70 75 80
Gly Ser Gly Phe Leu Leu Gly Ile Gly Ser Ala Cys Ala Ser Gly Val
85 90 95
Ala Val Ser Lys Val Leu His Leu Glu Gly Glu Val Asn Lys Ile Lys
100 105 110
Asn Ala Leu Leu Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly
115 120 125
Val Ser Val Leu Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp
130 135 140
Lys Glu Leu Leu Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn
145 150 155 160
Ile Ala Thr Val Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu
165 170 175
Ile Ala Arg Glu Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser
180 185 190
Thr Tyr Met Leu Thr Asn Ser Glu Leu Leu Ser Phe Ile Asn Asp Met
195 200 205
Pro Ile Thr Asn Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile
210 215 220
Val Arg Gln Gln Ser Tyr Ser Cys Met Ser Val Val Lys Glu Glu Val
225 230 235 240
Ile Ala Tyr Val Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro
245 250 255
Cys Trp Lys Leu His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu
260 265 270
Gly Ser Asn Ile Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp
275 280 285
Asn Ala Gly Ser Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val
290 295 300
Gln Cys Asn Arg Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro
305 310 315 320
Thr Asp Val Asn Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp
325 330 335
Cys Lys Ile Met Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr
340 345 350
Ser Ile Gly Ala Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser
355 360 365
Asn Lys Asn Arg Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr
370 375 380
Val Ser Asn Lys Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr
385 390 395 400
Tyr Val Asn Lys Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro
405 410 415
Ile Ile Asn Tyr Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp
420 425 430
Ala Ser Ile Ala Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe
435 440 445
Ile Arg Arg Ser Asp Glu Leu Leu His
450 455
<210> 11
<211> 453
<212> PRT
<213>Artificial sequence
<400> 11
Gln Asn Ile Thr Glu Glu Phe Tyr Gln Ser Thr Cys Ser Ala Val Ser
1 5 10 15
Arg Gly Tyr Leu Ser Ala Leu Arg Thr Gly Trp Tyr Thr Ser Val Val
20 25 30
Thr Ile Glu Leu Ser Lys Ile Gln Lys Asn Val Cys Lys Ser Thr Asp
35 40 45
Ser Lys Val Lys Leu Ile Lys Gln Glu Leu Glu Arg Tyr Asn Asn Ala
50 55 60
Val Val Glu Leu Gln Ser Leu Met Gln Cys Glu Pro Ala Ser Gly Ser
65 70 75 80
Gly Ser Gly Ile Gly Ser Ala Ile Ala Ser Gly Val Ala Val Ser Lys
85 90 95
Val Cys His Leu Glu Gly Glu Val Asn Lys Ile Lys Asn Ala Leu Leu
100 105 110
Ser Thr Asn Lys Ala Val Val Ser Leu Ser Asn Gly Val Ser Val Leu
115 120 125
Thr Ser Lys Val Leu Asp Leu Lys Asn Tyr Ile Asp Lys Glu Leu Leu
130 135 140
Pro Lys Val Asn Asn His Asp Cys Arg Ile Ser Asn Ile Ala Thr Val
145 150 155 160
Ile Glu Phe Gln Gln Lys Asn Asn Arg Leu Leu Glu Ile Ala Arg Glu
165 170 175
Phe Ser Val Asn Ala Gly Ile Thr Thr Pro Leu Ser Thr Tyr Met Leu
180 185 190
Thr Asn Ser Glu Leu Leu Ser Phe Ile Asn Asp Met Pro Ile Thr Asn
195 200 205
Asp Gln Lys Lys Leu Met Ser Ser Asn Val Gln Ile Val Arg Gln Gln
210 215 220
Ser Tyr Ser Ile Met Cys Val Val Lys Glu Glu Val Ile Ala Tyr Val
225 230 235 240
Val Gln Leu Pro Ile Tyr Gly Val Ile Asp Thr Pro Cys Trp Lys Leu
245 250 255
His Thr Ser Pro Leu Cys Thr Thr Asp Asn Lys Glu Gly Ser Asn Ile
260 265 270
Cys Leu Thr Arg Thr Asp Arg Gly Trp Tyr Cys Asp Asn Ala Gly Ser
275 280 285
Val Ser Phe Phe Pro Gln Ala Glu Thr Cys Lys Val Gln Cys Asn Arg
290 295 300
Val Phe Cys Asp Thr Met Asn Ser Leu Thr Leu Pro Thr Asp Val Asn
305 310 315 320
Leu Cys Asn Thr Asp Ile Phe Asn Thr Lys Tyr Asp Cys Lys Ile Met
325 330 335
Thr Ser Lys Thr Asp Ile Ser Ser Ser Val Ile Thr Ser Ile Gly Ala
340 345 350
Ile Val Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser Asn Lys Asn Arg
355 360 365
Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr Val Ser Asn Lys
370 375 380
Gly Val Asp Thr Val Ser Val Gly Asn Thr Leu Tyr Tyr Val Asn Lys
385 390 395 400
Leu Glu Gly Lys Ala Leu Tyr Ile Lys Gly Glu Pro Ile Ile Asn Tyr
405 410 415
Tyr Asp Pro Leu Val Phe Pro Ser Asp Glu Phe Asp Ala Ser Ile Ala
420 425 430
Gln Val Asn Ala Lys Ile Asn Gln Ser Leu Ala Phe Ile Arg Arg Ser
435 440 445
Asp Glu Leu Leu His
450

Claims (8)

1. a kind of bovine respiratory born of the same parents zoarium viral antigen proteins, encode the fit virus key protein F- albumen of bovine respiratory born of the same parents wild The aminoacid sequence that plants is abbreviation SEQ CAA76980.1, and particular sequence is MATTAMTMIISIIFISTYVTHITLCQNITEEFYQSTCSAVSRGYLSALRTGWYTSVVTIELSKIQKNVCKSTDSKVK LIKQELERYNNAVVELQSLMQNEPASFSRAKRSIPELIHYTRNSTKKFYGLMGKKRKRRFLGFLLGIGSAIASGVAV SKVLHLEGEVNKIKNALLSTNKAVVSLSNGVSVLTSKVLDLKNYIDKELLPKVNNHDCRISNIATVIEFQQKNNRLL EIAREFSVNAGITTPLSTYMLTNSELLSLINDMPITNDQKKLMSSNVQIVRQQSYSIMSVVKEEVIAYVVQLPIYGV IDTPCWKLHTSPLCTTDNKEGSNICLTRTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPTDVNLCNTD IFNTKYDCKIMTSKTDISSSVITSIGAIVCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTLYYVNKLE GKALYIKGEPIINYYDPLVFPSDEFDASIAQVNAKINQSLAFIRRSDELLHSVDVGKSTTNVVITTIIIVIVVVILM LIAVGLLFYCKTRSTPIMLGKDQLSGINNLSFSK, it is characterised in that：The bovine respiratory born of the same parents zoarium viral antigen proteins Aminoacid sequence compare at least two sites of SEQ CAA76980.1 and substituted by cysteine, and half Guang ammonia after substituting By disulfide bond between acid.
2. a kind of bovine respiratory born of the same parents zoarium viral antigen proteins according to claim 1, it is characterised in that：The site is The 148th L-Valine of aminoacid sequence and 288 isoleucine.
3. a kind of bovine respiratory born of the same parents zoarium viral antigen proteins according to claim 1, it is characterised in that：The site is The 158th Leucine of aminoacid sequence and 290 serines.
4. a kind of fit viral antigen proteins of bovine respiratory born of the same parents according to claim 1, it is characterised in that：The cattle exhales The aminoacid sequence for inhaling road born of the same parents zoarium viral antigen proteins compares SEQ CAA76980.1, the 260th site Leucine by half Cystine is substituted, and realizes hole filling in the site.
5. the fit viral antigen proteins of any one bovine respiratory born of the same parents according to claim 1 ~ 3, it is characterised in that：Described Aminoacid sequence for bovine respiratory born of the same parents zoarium viral antigen proteins rejects 103 sites in SEQ CAA76980.1 sequence To the aminoacid sequence in 138 sites, 102 sites and 139 sites are connected using sgsgs.
6. the fit viral antigen proteins of any one bovine respiratory born of the same parents according to claim 1 ~ 3, it is characterised in that：Described For 103 sites in the aminoacid sequence rejecting SEQ CAA76980.1 sequence of bovine respiratory born of the same parents zoarium viral antigen proteins extremely The aminoacid sequence in 142 sites, connects 102 sites and 143 sites using sgsgs.
7. the fit viral antigen proteins of any one bovine respiratory born of the same parents according to claim 1 ~ 3, it is characterised in that：Described The aminoacid sequence of coding bovine respiratory born of the same parents' zoarium viral antigen proteins compares SEQ CAA76980.1 sequence, the 99th site day Winter propylhomoserin is substituted by cysteine, and the 362nd site serine is substituted by cysteine, and is connected between the cysteine after replacement Connect interior disulfide bond.
8. the preparation method of a kind of bovine respiratory born of the same parents zoarium viral antigen proteins, it is characterised in that：Concrete technology is as follows：
（1）According to the principle of structure biology, protein crystal is carried out, observed with X-ray after crystallization, determine and infect virus key Cause of disease F- albumen infect before and after structure change；
（2）According to step（1）Structure change designs the sequence of antigen protein and structure；
（3）Determine and need the qualification of antigen protein through multiple antigen tests and stability characteristic detection；
（4）Antigen protein sequence through checking is carried out corresponding albumen synthesis, expression, purification and is detected, obtain for making For animal experiment and clinical trial for bovine respiratory born of the same parents zoarium viral antigen proteins.
CN201611065849.2A 2016-11-28 2016-11-28 Antigen protein of bovine respiration syncytial viruses Pending CN106432437A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1264425A (en) * 1997-07-17 2000-08-23 皮埃尔法博赫药品公司 Syncytial respiratory virus epitopes and antibodies comprising them, useful in diagnosis and therapy
CN103842374A (en) * 2011-05-13 2014-06-04 诺华股份有限公司 Pre-fusion rsv f antigens

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1264425A (en) * 1997-07-17 2000-08-23 皮埃尔法博赫药品公司 Syncytial respiratory virus epitopes and antibodies comprising them, useful in diagnosis and therapy
CN103842374A (en) * 2011-05-13 2014-06-04 诺华股份有限公司 Pre-fusion rsv f antigens

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BRUCE,C.J.: "F protein [Bovine orthopneumovirus]", 《GENBANK DATABSE》 *

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